The present invention relates to a bioreactor apparatus, and particularly relates to a bioreactor apparatus for producing metabolites such as human body cure vaccines, interferons, cancer antigens, hormones, cell growth factors, lymphocines, various kinds of catalysts, and the like, from bioorganisms such as various kinds of animal cells and the like through depositing the animal cells on a carrier made of a porous material such as nonwoven fabric or the like, and cultivating the animal cells with a high yield and density on the carrier, by which various useful physiologically active substances can be produced selectively, continuously and efficiently.
For in-vitro production of useful substances produced by various human cells, such as human immuno chemical mediators, human type enzymes, etc., for medicinal application it, is necessary to establish both a technique for mass-cultivating various kinds of animal cells and develop a high-performance bioreactor apparatus.
The animal cell cultivating techniques used heretofore include "a suspension growth technique" for growing cells suspended in an aqueous solution, "a microcarrier growth technique" for growing suspended cells by adhering the cells with porous microbeads having a diameter of about 200 .mu.m, "an adhesion carrier growth technique" for growing cells by carrying the cells on an adequate nonporous and/or porous carrier (such as porous film, hollow fiber, nonwoven fabrics etc), and the like.
Japanese Patent Unexamined Publication No. Sho. 59-59187 discloses a cultivating apparatus for performing the adhesion carrier growth technique in which animal cells are adhered or grown on various forms of culture supports housed in a culture container of the apparatus while a culture solution is sprayed and supplied into the container.
Japanese Patent Unexamined Publication No. Sho. 56-42584 discloses a cultivating apparatus designed to pack hollow fibers in a culture container fill space between a wall of the container and an outer surface of the hollow fiber with suspended cells and supply a culture solution from the inside of the hollow fiber. Further, Japanese Patent Unexamined Publication No. Sho. 64-34276 discloses a material for performing the adhesion carrier growth technique, in which nonwoven fabric made from specific superfine fiber is used as a carrier for a cell culture bed and is arranged in a plastic schale (laboratory dish) to cultivate animal cells.
However, in the conventional suspension growth technique and the microcarrier growth technique, the concentration of floated or suspended animal cells is limited to a maximum of about 10 g/l. Further, the concentration of the substrate solution cannot be increased because of substrate inhibition and osmotic pressure. Furthermore, shear forces imposed on cells in the solution by contact, collision, abrasion, etc., of the cells with the fluidal change of solvent are so large that the cells are often inactivated or killed. In the microcarrier growth technique, the cells are, in most cases, separated or dropped from the microbeads. In the adhesion carrier growth technique, it is not only difficult to continuously circulate and exchange the culture solution and the substrate solution and to supply the necessary oxygen for cultivating animal cells, but the contact of the cells with the culture solution and the substrate solution is uneven, insufficient and inefficient. Furthermore, metabolites cannot be continuously separated to the outside of the apparatus. As a result, nutritional balance disorder caused by excess or shortage of the substrate and culture environment disorder caused by storage of the metabolites occur locally in the cells. Consequently, animal cells cannot be cultivated in a high yield and a high density per volume. Further, efficient metabolite production is impossible. Among the conventional apparatuses for performing the aforementioned techniques, there is no apparatus for solving these problems.